Acid Lipase Activity Research Articles

From LAL-D to MASLD: Insights into the role of LAL and Kupffer cells in liver inflammation and lipid metabolism

Metabolic dysfunction-associated steatotic liver disease (MASLD) is a prevalent liver pathology worldwide, closely associated with obesity and metabolic disorders. Increasing evidence suggests that macrophages play a crucial role in the development of MASLD. Several human studies have shown an inverse correlation between circulating lysosomal acid lipase (LAL) activity and MASLD. LAL is the sole enzyme known to degrade cholesteryl esters (CE) and triacylglycerols in lysosomes. Consequently, these substrates accumulate when their enzymatic degradation is impaired due to LAL deficiency (LALD). This study aimed to investigate the role of hepatic LAL activity and liver-resident macrophages (i.e., Kupffer cells (KC)) in MASLD. To this end, we analyzed lipid metabolism in hepatocyte-specific (hep)Lal−/− mice and depleted KC with clodronate treatment. When fed a high-fat/high-cholesterol diet (HF/HCD), hepLal−/− mice exhibited CE accumulation and an increased number of macrophages in the liver and significant hepatic inflammation. KC were depleted upon clodronate administration, whereas infiltrating/proliferating CD68-expressing macrophages were less affected. This led to exacerbated hepatic CE accumulation and dyslipidemia, as evidenced by increased LDL-cholesterol concentrations. Along with proteomic analysis of liver tissue, these findings indicate that hepatic LAL-D in HF/HCD-fed mice leads to macrophage infiltration into the liver and that KC depletion further exacerbates hepatic CE concentrations and dyslipidemia.

An Investigation of Volatile Flavor Compounds and Lipolysis-Oxidation in Coppa as Affected by the Inoculation of Coagulase-Negative Staphylococcus during the Air-Drying Stage.

This study aimed to explore the effects of coagulase-negative Staphylococcus inoculation on flavor generation and lipolysis-oxidation in Coppa. Acid lipase, neutral lipase, phospholipase, and lipoxygenase (LOX) activities, as well as free fatty acids, volatile compounds, and sensory evaluation, were determined during the fermentation and air-drying processes of Coppa over 40 days. Staphylococcus carnosus and Staphylococcus xylosus or a combination of both strains were selected for this study, and natural fermentation was treated as a control. The results showed that Staphylococcus inoculation significantly enhanced lipase and LOX activities, and mixed strains had a superior effect. Palmitic acid, stearic acid, linoleic acid, and oleic acid were identified as the predominant free fatty acids in Coppa, with the mixed fermentation group exhibiting the highest contents. Acids, aldehydes, alcohols, ketones, esters, and phenols were found for the volatile compounds in Coppa. These findings thus suggested a positive role of Staphylococcus inoculation in activating lipolysis-oxidation and contributing to the flavor formation of Coppa during the air-drying stage.

Cholesterol-rich lysosomes induced by respiratory syncytial virus promote viral replication by blocking autophagy flux

Respiratory syncytial virus (RSV) hijacks cholesterol or autophagy pathways to facilitate optimal replication. However, our understanding of the associated molecular mechanisms remains limited. Here, we show that RSV infection blocks cholesterol transport from lysosomes to the endoplasmic reticulum by downregulating the activity of lysosomal acid lipase, activates the SREBP2–LDLR axis, and promotes uptake and accumulation of exogenous cholesterol in lysosomes. High cholesterol levels impair the VAP-A-binding activity of ORP1L and promote the recruitment of dynein–dynactin, PLEKHM1, or HOPS VPS39 to Rab7–RILP, thereby facilitating minus-end transport of autophagosomes and autolysosome formation. Acidification inhibition and dysfunction of cholesterol-rich lysosomes impair autophagy flux by inhibiting autolysosome degradation, which promotes the accumulation of RSV fusion protein. RSV-F storage is nearly abolished after cholesterol depletion or knockdown of LDLR. Most importantly, the knockout of LDLR effectively inhibits RSV infection in vivo. These findings elucidate the molecular mechanism of how RSV co-regulates lysosomal cholesterol reprogramming and autophagy and reveal LDLR as a novel target for anti-RSV drug development.

Frequency of rs1051338 and rs116928232 Variants in Individuals from Northwest Mexico.

LIPA, situated on chromosome 10q23.2-q23.3, encodes the enzyme lysosomal acid lipase (LAL) (EC 3.1.1.13). Genetic alterations in LIPA lead to lysosomal acid lipase deficiency (LALD), an inborn error causing lipid metabolism anomalies and impairing cholesterol and triacylglyceride degradation. Over 40 LIPA variants have been documented, yet this study focuses on just two. The rs1051338 variant (NM_000235:c.46A>C) affects the signal peptide in Exon 2, whereas rs116928232, located in Exon 8, alters the splice site (NM_000235:c.894G>A), impacting lysosomal acid lipase activity. Considering the diverse clinical manifestations of LALD and the rising hepatic steatosis prevalence in Mexican population, mainlydue to diet, these variants were investigated within this demographic to uncover potential contributing factors. This study aimed to reveal the frequency of rs1051338 and rs116928232 among healthy mestizo individuals in Northwest Mexico, marking a significant genetic exploration in this demographic. Three hundred ten healthy mestizo individuals underwent PCR-RFLP analysis for both variants, and Sanger sequencing was performed for variant rs116928232. Bioinformatic analysis was also performed to predict protein changes. Allele frequencies for rs1051338 (FA = 0.39, p value = 0.15) and rs116928232 (FA = 0.0016, p value = 0.49) aligned with reported data, while bioinformatic analysis allowed us to identify the protein alteration observed in both variants; finally, the variants showed no linkage between them (normalized D' = 1.03, p value = 0.56). Allelic frequencies closely matched reported data, and protein structure analysis confirmed variant impacts on LAL enzyme function. Notably, this study marks the first analysis of rs1051338 and rs116928232 in a healthy Mexican mestizo population.

Abstract 1061: The Phospholipase cγ2 And Protein Kinase cα Signaling Axis Is A Key Regulator Of The Extracellular Digestion Of Lipoprotein Aggregates By Macrophages

Atherosclerosis is characterized by cholesterol ester accumulation in macrophages, leading to foam cell formation. In atherosclerotic lesions, most lipoproteins are aggregates (agLDL) that are tightly crosslinked to the extracellular matrix, making digesting them by endocytosis or phagocytosis difficult. Thus, it has been unclear how the cholesteryl esters in the core of agLDL are hydrolyzed. Our laboratory discovered a novel mechanism macrophages use to digest agLDL and generate free cholesterol in an extracellular, acidic, hydrolytic compartment known as the lysosomal synapse. Macrophages form a tight seal around agLDL through actin polymerization and deliver lysosomal contents into this space in a process termed digestive exophagy. Vacuolar ATPase on the plasma membrane lowers the pH of the lysosomal synapse, enabling lysosomal acid lipase (LAL) activity. LAL generates free cholesterol, which accumulates in macrophages and leads to foam cell formation. Our laboratory has begun to characterize the signaling pathways that regulate digestive exophagy, having previously identified TLR4 activation of MyD88/Syk as critical. Here, our goal was to identify digestive exophagy regulators downstream of Syk. Syk activates Bruton’s tyrosine kinase (BTK) and phospholipase Cγ2 (PLCγ2). We tested the hypothesis that Syk phosphorylates and activates PLCγ2 during digestive exophagy. Using pharmacological treatment and genetic manipulation, we found that PLCγ2 and to a lesser extent BTK regulate digestive exophagy. We confirmed that inhibition of Syk or BTK leads to a loss of PLCγ2 activation. PLCγ2 cleaves PI(4,5)P 2 into diacylglycerol (DAG) and inositol 1,4,5-trisphosphate (IP 3 ). Soluble IP 3 then activates the release of Ca 2+ from the endoplasmic reticulum (ER) into the cytoplasm. We demonstrated that Ca 2+ release from the ER is upregulated by agLDL and plays a role in digestive exophagy. Both DAG and Ca 2+ activate protein kinase Cα (PKCα). We also found that PKCα regulates digestive exophagy. In conclusion, we identified new signaling mechanisms regulating digestive exophagy downstream of Syk. We hope that expanding our understanding of the mechanisms of digestive exophagy will be helpful in identifying therapeutics to slow atherosclerosis.

Pedigree Analysis of Nonclassical Cholesteryl Ester Storage Disease with Dominant Inheritance in a LIPA I378T Heterozygous Carrier.

Cholesterol ester storage disorder (CESD; OMIM: 278,000) was formerly assumed to be an autosomal recessive allelic genetic condition connected to diminished lysosomal acid lipase (LAL) activity due to LIPA gene abnormalities. CESD is characterized by abnormal liver function and lipid metabolism, and in severe cases, liver failure can occur leading to death. In this study, one Chinese nonclassical CESD pedigree with dominant inheritance was phenotyped and analyzed for the corresponding gene alterations. Seven males and eight females from nonclassical CESD pedigree were recruited. Clinical features and LAL activities were documented. Whole genome Next-generation sequencing(NGS) was used to screen candidate genes and mutations, Sanger sequencing confirmed predicted mutations, and qPCR detected LIPA mRNA expression. Eight individuals of the pedigree were speculatively thought to have CESD. LAL activity was discovered to be lowered in four living members of the pedigree, but undetectable in the other four deceased members who died of probable hepatic failure. Three of the four living relatives had abnormal lipid metabolism and all four had liver dysfunctions. By liver biopsy, the proband exhibited diffuse vesicular fatty changes in noticeably enlarged hepatocytes and Kupffer cell hyperplasia. Surprisingly, only a newly discovered heterozygous mutation, c.1133T>C (p. Ile378Thr) on LIPA, was found by gene sequencing in the proband. All living family members who carried the p.I378T variant displayed reduced LAL activity. Phenotypic analyses indicate that this may be an autosomal dominant nonclassical CESD pedigree with a LIPA gene mutation.

S1423 Serum Lysosomal Acid Lipase Activity as a Potential Non-Invasive Diagnostic Marker for Non-Alcoholic Fatty Liver Disease: A Systematic Review and Meta-Analysis

S1423 Serum Lysosomal Acid Lipase Activity as a Potential Non-Invasive Diagnostic Marker for Non-Alcoholic Fatty Liver Disease: A Systematic Review and Meta-Analysis

Effects of low salt on lipid oxidation and hydrolysis, fatty acids composition and volatiles flavor compounds of dry-cured ham during ripening

In order to study the effect of low salt on lipid oxidation and volatile flavor of dry-cured ham, a formula containing 18% potassium lactate, 12% lysine and 70% NaCl was used as an alternative salt for curing ham. The control group using 100% NaCl as pickled formulation dry-cured ham.The results showed that low salt significantly increased peroxide value (POV) and thiobarbituric acid value (TBARS) (P < 0.05). Acid lipase, neutral lipase, and phospholipase enzyme activity, as well as the amount of free fatty acids (FFAs), were all considerably higher in the low salt group than in the full salt group (P < 0.05). The volatile flavor compounds of dry-cured ham were determined by GC-MS, and 14 differential metabolites were screened out by using OPLS-DA analysis based on P < 0.05 and VIP >1. The enrichment analysis showed that low salt mainly affected low molecules fatty acids (palmitic acid, stearic acid, linoleic acid and myristic acid). Meanwhile, low salt treatment also influenced biosynthesis of unsaturated fatty acids, linoleic acid metabolism, fatty acid biosynthesis, fatty acid elongation and fatty acid degradation. This study demonstrates that low salt promotes the formation of ham flavor quality by affecting lipid oxidation and hydrolysis.

Is lysosomal acid lipase activity associated with the presence and severity of coronary artery disease?

Lipid metabolism is considerably complex and there can be many critical steps in atherogenesis. The association between lysosomal acid lipase (LAL) activity and coronary artery disease (CAD) has not been elucidated in detail. We aimed to evaluate the association between LAL activity with the presence and severity of CAD in patients who are seen in daily clinical practice. Patients who underwent coronary angiography were divided into groups according to the angiography results. Syntax scores and Gensini scores were calculated. The LAL activity was measured from dried blood spots. Median LAL activity values were similar in all study groups (normal coronary arteries: 0.40 nmol/punch/h; non-obstructive CAD: 0.44 nmol/punch/h; obstructive chronic CAD: 0.40 nmol/punch/h; obstructive acute coronary syndrome: 0.48 nmol/punch/h) and there was no correlation between coronary atherosclerotic burden and LAL activity (correlation coefficients Syntax score and LAL: -0.032; Gensini score and LAL: -0.030). In addition, no relationship between serum lipid levels and LAL activity was detected. The presence of CAD and its severity is not associated with the LAL activity in patients encountered in daily clinical practice.

Bran from composite grain mixture as an object of deep processing. Part 2. Carbohydrate-amylase and lipid complexes

Polycomponent bran obtained by joint grinding of a grain mixture from cereals (wheat), legumes (lentils) and oilseeds (flax) should be considered as a valuable secondary raw material, the use of which for deep processing will make it possible to obtain various food and feed ingredients. An assessment of the carbohydrate-amylase and lipid complexes of the three variants of multicomponent bran showed that the ratio of amylose and amylopectin in the starch of three-component bran is almost the same, however, in lentil-flax bran, the proportion of amylose is 1.6 times higher than in the first two variants. The specific activity of amylases in three-component bran is about 2 times higher than in lentil-flax bran. In addition, the latter are characterized by a higher content of reducing sugars and fiber. The molecular weight of amylases isolated from three-component bran, according to gel chromatography, was: α-amylase — 40,000 Da; β-amylase — 60,000 Da. It has been established that the addition of flax seeds to the grinding mixture significantly increases the fat content in bran 6.4; 6.0 and 12.9%. The fatty acid composition of the studied bristles is characterized by the predominance of unsaturated fatty acids. At the same time, the ratio of essential acids — linoleic acid (ɷ‑6) to α-linolenic acid (ɷ‑3) in favor of the most deficient α-linolenic acid — was typical for lentil-flax bran and amounted to 1:4.2. The activity of alkaline lipases, which exhibit their effect at pH 8.0 (mainly cereal lipases), and acid lipases (mainly oilseed lipases) with an optimum of action at pH 4.7 in three-component bran samples are approximately the same, and lentil-flax bran is characterized by a high specific acid lipase activity, which is approximately 4.2 times higher than the activity of acid lipases of three-component bran. The data obtained, along with data on the characteristics of the protein-proteinase complex of the studied types of bran, will be used in the development of methods for enzymatic modification (deep processing) and in the preparation of components for the creation of new food products with increased nutritional and biological value.

Lipase‐a single‐nucleotide polymorphism rs143793106 is associated with increased risk of aggressive periodontitis by negative influence on the cytodifferentiation of human periodontal ligament cells

Aggressive periodontitis (AgP) is characterized by general health and rapid destruction of periodontal tissue. The familial aggregation of this disease highlights the involvement of genetic factors in its pathogeny. We conducted a genome-wide association study (GWAS) to identify AgP-related genes in a Japanese population, and the lipid metabolism-related gene, lipase-a, lysosomal acid type (LIPA), was suggested as an AgP candidate gene. However, there is no report about the expression and function(s) of LIPA in periodontal tissue. Hence, we studied the involvement of how LIPA and its single-nucleotide polymorphism (SNP) rs143793106 in AgP by functional analyses of LIPA and its SNP in human periodontal ligament (HPDL) cells. GWAS was performed using the genome database of Japanese AgP patients, and the GWAS result was confirmed using Sanger sequencing. We examined the mRNA expression level of LIPA and the protein expression level of the encoded protein lysosomal acid lipase (LAL) in periodontium-composing cells using conventional and real-time polymerase chain reaction (PCR) and western blotting, respectively. Lentiviral vectors expressing LIPA wild-type (LIPA WT) and LIPA SNP rs143793106 (LIPA mut) were transfected into HPDL cells. Western blotting was performed to confirm the transfection. LAL activity of transfected HPDL cells was determined using the lysosomal acid lipase activity assay. Transfected HPDL cells were cultured in mineralization medium. During the cytodifferentiation of transfected HPDL cells, mRNA expression of calcification-related genes, alkaline phosphatase (ALPase) activity and calcified nodule formation were assessed using real-time PCR, ALPase assay, and alizarin red staining, respectively. The GWAS study identified 11 AgP-related candidate genes, including LIPA SNP rs143793106. The minor allele frequency of LIPA SNP rs143793106 in AgP patients was higher than that in healthy subjects. LIPA mRNA and LAL protein were expressed in HPDL cells; furthermore, they upregulated the cytodifferentiation of HPDL cells. LAL activity was lower in LIPA SNP-transfected HPDL cells during cytodifferentiation than that in LIPA WT-transfected HPDL cells. In addition, ALPase activity, calcified nodule formation, and calcification-related gene expression levels were lower during cytodifferentiation in LIPA SNP-transfected HPDL cells than those in LIPA WT-transfected HPDL cells. LIPA, identified as an AgP-related gene in a Japanese population, is expressed in HPDL cells and is involved in regulating cytodifferentiation of HPDL cells. LIPA SNP rs143793106 suppressed cytodifferentiation of HPDL cells by decreasing LAL activity, thereby contributing to the development of AgP.

Effect of Dietary Lipid Level on Growth Performance, Body Composition, and Physiometabolic Responses of Genetically Improved Farmed Tilapia (GIFT) Juveniles Reared in Inland Ground Saline Water.

A 60-day feeding trial was carried out to determine the effect of dietary lipid levels on growth and physiometabolic responses to optimize the dietary lipid requirement for maximizing the growth of Genetically Improved FarmedTilapia (GIFT) juveniles reared in inland ground saline water (IGSW) of medium salinity (15 ppt). Formulation and preparation of seven heterocaloric (389.56-449.02 Kcal digestible energy/100 g), heterolipidic (40-160 g/kg), and isonitrogenous (410 g/kg crude protein) purified diets were done for conducting the feeding trial. Random distribution of 315 acclimatized fish (mean weight 1.90 ± 0.01 g) was made in seven experimental groups such as CL4 (40 g/kg lipid), CL6 (60 g/kg lipid), CL8 (80 g/kg lipid), CL10 (100 g/kg lipid), CL12 (120 g/kg lipid), CP14 (140 g/kg lipid), and CL16 (160 g/kg lipid) with 15 fish per triplicate tank (fish density, 0.21 kg/m3). Respective diets were used for feeding the fish at satiation level three times daily. Results indicated that weight gain percentage (WG%), specific growth rate (SGR), protein efficiency ratio, and protease activity significantly increased up to 100 g lipid/kg fed group, and then the values significantly decreased. Muscle ribonucleic acid (RNA) content and lipase activity were highest in 120 g/kg lipid-fed group. RNA/DNA (deoxyribonucleic acid) and serum high-density lipoproteins levels of 100 g/kg lipid-fed group were significantly higher than 140, and 160 g/kg lipid-fed groups. The lowest feed conversion ratio was found in the 100 g/kg lipid-fed group. The amylase activity was significantly higher in 40 and 60 g lipid/kg fed groups. The whole-body lipid level was increased with increasing the dietary lipid levels, whereas, there was no significant difference in whole-body moisture, crude protein, and crude ash contents of all groups. Highest serum glucose, total protein and albumin, and albumin to globulin ratio and lowest low-density lipoproteins level were found in 140 and 160 g/kg lipid-fed groups. Serum osmolality and osmoregulatory capacity did not vary significantly, whereas carnitine palmitoyltransferase-I and glucose-6-phosphate dehydrogenase showed an increased and decreased trend, respectively, with the increasing dietary lipid levels. According to second-order polynomial regression analysis based on WG% and SGR, the optimum dietary lipid for GIFT juveniles in IGSW of 15 ppt salinity was found to be 99.1 and 100.1 g/kg, respectively.

Effect of sodium replacement on the quality characteristics of pastırma (a dry-cured meat product)

This study aims to investigate the effect of sodium replacement on the quality characteristics of pastırma. For this purpose, pastırma production with four different salt mixtures (I. 100 % NaCl; II. 50 % NaCl + 50 % KCl; III. 40 % NaCl + 40 % KCl + 20 % CaCl2; IV. 30 % NaCl + 40 % KCl + 20 % CaCl2 + 10 % MgCl2) were carried out using traditional method. The use of different salt mixtures for pastırma had no statistically significant effect on the microbial counts and residual nitrite of the final product. The a* and b* values were affected by this treatment. The salt mixture containing CaCl2 or CaCl2 + MgCl2 significantly decreased the pH values and sensory scores. The eighteen volatile compounds were affected by the salt mixtures. Na+, K+, Ca2+, and Mg2+ had a certain increase depending on their ratio in the salt mixtures. The highest Cathepsin activity in all pastırma samples was observed in Cathepsin B + L. The salt mixture with NaCl + KCl + CaCl2 + MgCl2 increased acid lipase activity. However, this mixture had no significant effect on neutral lipase activity.

Marine sources for biotechnology: Preliminary digestive enzymes assessment of seven decapod species from the southwest Atlantic

Marine natural compounds are required by industries to improve and reduce costs of many biotechnological processes. In this work, specific activity of midgut gland digestive enzymes was measured in seven species of decapods. Alkaline and acid peptidase, lipase and amylase activities were registered in Pleoticus muelleri (Pm), Artemesia longinaris (Al), Paralomis granulosa (Pg), Lithodes santolla (Ls), Munida gregaria (Mg), Neohelice granulata (Ng), and Ovalipes trimaculatus (Ot). The highest activities for all the analyzed digestive enzymes were obtained in Pg, Ls, Mg and Ot, which were used for optimum temperature and pH measurements and to test their stability at different temperatures and pH. In general, optimal enzyme temperatures were between 30 and 50 °C in the four crustaceans. Peptidases and lipases from these species showed in general alkaline optimal pH except for those from Ot which presented high activity at the whole pH range (2–12). Amylase from Ot presented its optimal at pH 6–8 and 30–50 °C. Thermal stability of peptidases was elevated at 30 °C for the four species tested (Pg; Ls; Mg and Ot), lipases were stable mainly at 30–50 °C and amylase was stable at 10 °C, 30 °C and 50 °C during the first 30 min of the assay. In general, stability of the four enzymes from Pg was greater at alkaline pH than at acid. The other species showed stability at all pH for the four enzymes. The range of temperature and pH of the tested enzymes activity are coincident with those used in various biotechnological processes and therefore indicates the potential of these enzymes to be used for industrial procedures. Further, most of the species studied are converted into fishery residues, either by processing or by-catch, so their reuse would also contribute to the sustainability of the activity.

RWD116 Reduced Lysosomal Acid Lipase Activity in Non-Alcoholic Fatty Liver Disease: A Systematic Review of Current Evidence and Future Directions

Non-alcoholic fatty liver disease (NAFLD) presents with accumulation of excessive intra-hepatic fat and acts as a major health burden globally. A multifactorial pathogenesis is reported to be involved. Reduced Lysosomal Acid Lipase (LAL) activity is suggested as one of the involved pathogenic mechanisms. This review summarizes the available evidence on role of reduced LAL activity in the pathogenesis of NAFLD.

Reduced lysosomal acid lipase (LAL) activity in NAFLD is related to the severity of liver disease ‘‘First study from the Asia-Pacific’’

Background and Aim: Non-alcoholic fatty liver disease (NAFLD) is estimated to be the most common liver disorder and presents as a major health burden globally. Pathogenesis of NAFLD is multi-factorial. Lysosomal Acid Lipase impairment is suggested as one of the involved pathogenic mechanisms in NAFLD. This study aimed to explore LAL activity in patients with NAFLD and its relationship with the disease severity.

Effects of Dietary Fermented Soybean Meals on Tissue Lipid Level, Bile Acid Concentration, Lipase Activity and Lipid Digestibility in Pompano Fish (Trachinotus blochii)

Effects of Dietary Fermented Soybean Meals on Tissue Lipid Level, Bile Acid Concentration, Lipase Activity and Lipid Digestibility in Pompano Fish (Trachinotus blochii)

Molecular and histological traits of reduced lysosomal acid lipase activity in the fatty liver

Recent studies demonstrated reduced blood lysosomal acid lipase (LAL) activity in patients with nonalcoholic fatty liver disease (NAFLD). We aimed to verify hepatic LAL protein content and activity in in vitro and in vivo models of fat overload and in NAFLD patients. LAL protein content and activity were firstly evaluated in Huh7 cells exposed to high-glucose/high-lipid (HGHL) medium and in the liver of C57BL/6 mice fed with high-fat diet (HFD) for 4 and 8 months. LAL protein was also evaluated by immunohistochemistry in liver biopsies from 87 NAFLD patients and 10 controls, and correlated with hepatic histology. Huh7 cells treated with HGHL medium showed a significant reduction of LAL activity, which was consistent with reduced LAL protein levels by western blotting using an antibody towards the N-term of the enzyme. Conversely, antibodies towards the C-term of the enzyme evidenced LAL accumulation, suggesting a post-translational modification that masks the LAL N-term epitope and affects enzymatic activity. Indeed, we found a high rate of ubiquitination and extra-lysosomal localization of LAL protein in cells treated with HGHL medium. Consistent with these findings, inhibition of proteasome triggered dysfunctional LAL accumulation and affected LAL activity. Accumulation of ubiquitinated/dysfunctional LAL was also found in the liver of HFD fed mice. In NAFLD patients, hepatic levels of non-ubiquitinated/functional LAL were lower than in controls and inversely correlated with disease activity and some of the hallmarks of reduced LAL. Fat overload leads to LAL ubiquitination and impairs its function, possibly reducing hepatic fat disposal and promoting NAFLD activity.

Associations between persistent organic pollutants and endometriosis: A multiblock approach integrating metabolic and cytokine profiling

Humans are exposed daily to complex mixtures of chemical pollutants through their environment and diet, some of which have the potential to disrupt the bodies’ natural endocrine functions and contribute to reproductive diseases like endometriosis. Increasing epidemiological and experimental evidence supports the association between endometriosis and certain persistent organic pollutants (POPs) like dioxins; however, little is known about the underlying linking mechanisms. The main objective of this study is to proof the methodological applicability and discovery potential of integrating ultra-trace mass spectrometry (MS) profiling of POP biomarkers and endogenous biomarker profiling (MS metabolomics and cytokines) in a case-control study for the etiological research of endometriosis. The approach is applied in a pilot clinical-based study conducted in France where women with and without surgically confirmed endometriosis were recruited. Serum samples were analysed with high-resolution MS for about 30 polychlorinated biphenyls (PCBs), organochlorinated pesticides and perfluoroalkyl substances (PFAS). About 600 serum metabolites and lipids were identified with targeted metabolomics using tandem MS with the Biocrates MxP® Quant 500 Kit. A panel of 4 pro-inflammatory cytokines were analysed using ELISA-based 4-PLEX analyser. Statistical analysis included a battery of variable selection approaches, multivariate logistic regression for single-chemical associations, Bayesian kernel machine regressions (BKMR) to identify mixture effects of POPs and a multiblock approach to identify shared biomarker signatures among high risk clusters. The results showed the positive associations between some POPs and endometriosis risk, including the pesticide trans-nonachlor Odds Ratio (95% Confidence Interval) 3.38 (2.06–5.98), p < 0.0001 and PCB 114 OR (95% CI) 1.83 (1.17–2.93), p = 0.009. The BKMR approach showed a tendency of a positive cumulative effect of the mixture, however trans-nonachlor exhibited significant associations within the mixture and interacted with other PCBs, strengthening the effects at highest concentrations. Finally, the multiblock analysis, relating the various blocks of data, revealed a latent cluster of women with higher risk of endometrioma presenting higher concentrations of trans-nonachlor, PCB 114 and dioxin-like toxic equivalents from PCBs, together with an increased inflammatory profile (i.e. elevated interleukin-8 and monocyte chemoattractant protein-1). It was also highlighted a specific metabolic pattern characterized by dysregulation of bile acid homeostasis and lipase activity. Further research will be required with larger sample size to confirm these findings and gain insight on the underlying mechanisms between POPs and endometriosis.

Effects of the addition of exogenous lipase on lipolysis and lipid oxidation during wet‐curing and dry‐ripening of silver carp inoculated with mixed starter cultures

SummaryLipid degradation caused by lipolysis and oxidation plays an important role in the evolution of final product quality of cured silver carp. This study aimed to determine the impact of exogenous lipase (ExL) on lipolysis and lipid oxidation during the wet‐curing and dry‐ripening of silver carp inoculated with mixed starter cultures. The addition of ExL to silver carp increased the hydrolysis of total lipids and phospholipids. The free fatty acids (FFAs) content during wet‐curing was significantly higher in the ExL‐treated group compared with the control. Increased FFA accumulation contributed to a high level of conjugated dienes, peroxides and thiobarbituric acid‐reactive substances in the ExL‐treated group, indicating that the lipid oxidation was improved by the addition of ExL. Increased neutral lipase, acid lipase and phospholipase activities during the wet‐curing stage might explain the more extensive release of FFAs in the presence of ExL. The addition of ExL promotes the hydrolysis and oxidation of lipids in cured silver carp.